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The lymphocyte growth effect in the supernatants from k 562 cell culture on healthy human peripheral blood lymphocytes

, : The lymphocyte growth effect in the supernatants from k 562 cell culture on healthy human peripheral blood lymphocytes. Journal of Catholic Medical College 38(1): 93-100

Although T cell growth factor (TCGF) is present in the conditioned medium from human T lymphocytes, the concentration of TCGF in the supernatants is very low and the activity is inconsistent. Instead of normal human T lymphocytes, to obtain the lymphocyte growth effect from other cell line, myelogenous leukemic cell, K562 was cultured at a cell concentration of 1 .times. 106/ml in 4 different conditioned media and each supernatant was harvested. TCGF activity was assessed by the addition of K562 supernatants to 3 or 12 days cultured normal human lymphocytes prestimualted with 1% PHA-M [phytohemagglutinin]. The test cultures were incubated at C, 5% CO2, air flow for 48 h and pulsed with 3H-thymidine for 6 h before harvesting. Supernatants for K562 cells cultured with RPMI-1640 media containing 10% fetal calf serum had higher effect on the growth of lymphocytes than those cultured with RPMI-1640 media containing no fetal calf serum, but they did not show increased effect by the addition of 1% PHA-M or 2 .mu.g/ml Con A [concanavalin A] to RPMI-1640 meida containing 10% fetal calf serum. K562 cells were cultured for 1, 2, 3, 4 and 5 days and the supernatants from K562 cells cultured for 3 days showed the highest lymphocyte growth effect. Assessment by lymphocytes cultured with 1% PHA-M showed that the lymphocytes growth effect of supernatants from K562 cell culture was higher when lymphocytes were cultured for 12 days than 3 days. The lymphocyte growth effect was maintained high (stimulation index 10.3) until 32:1 dilution when supernatants from K562 cells cultured for 3 days with RPMI-1640 media containing 10% fetal calf serum continued to be diluted 2 fold.

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